The purpose of this study was to investigate the diffusion of calcium ions (Ca+2) through exposed dentinal tubules following intracanal application of mineral trioxide aggregate (MTA). Fifty-two single-rooted teeth were instrumented using 2.5% sodium hypochlorite for irrigation between each file size. Thereafter, standardized defects were created on the root surfaces so as to mimic external root resorption. The root canals and external defects received a final irrigation of 17% ethylenediaminetetraacetic acid and distilled water. MTA powder was then mixed with saline and placed into the canals. All root surfaces except the cavities were sealed with two coats of varnish. Teeth with unfilled canals (n = 26) served as controls. The teeth were immersed in saline after which the release of Ca+2 from the defects into the saline was measured at 1, 3, 7, 14, and 28 days. The results showed diffusion of Ca+2 through the defects in the dentin in MTA-filled roots with a significant increase in concentration within time.